Daniel:Notebook/HiResChrPaint/2013-5-11: Difference between revisions

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==DpnII Digestion==
==DpnII Digestion==


[[Daniel:Reagents/DpnII|Info]]
[[Daniel:Reagents/DpnII|Enzyme Info]]


  1. Prepared samples according to table
  1. Prepared samples according to table
  *DpnII-Lot 0091201; Buffer 0031105
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext>
|- style="font-size:12pt;font-weight:bold" align="center"
| width="65" height="40" | Sample
| width="65" | V4A
| width="65" | V4B
| width="65" | V4C
| width="65" | V4D
| width="65" | V6A
| width="65" | V6B
| width="65" | V6C
| width="65" | V6D
|- style="font-size:12pt" align="center"
|style="font-weight:bold" height="15" | ng/uL
| align="center" | 127.5
| align="center" | 156.3
| align="center" | 130.2
| align="center" | 130.6
| align="center" | 142.7
| align="center" | 138.9
| align="center" | 146.4
| align="center" | 133.4
|- style="font-size:12pt" align="center"
|style="font-weight:bold" height="15" | ng
| align="center" | 5100
| align="center" | 6252
| align="center" | 5208
| align="center" | 5224
| align="center" | 5708
| align="center" | 5556
| align="center" | 5856
| align="center" | 5336
|- style="font-size:12pt" align="center"
|style="font-weight:bold" height="15" | uL
| align="center" | 35
| align="center" | 35
| align="center" | 35
| align="center" | 35
| align="center" | 35
| align="center" | 35
| align="center" | 35
| align="center" | 35
|- style="font-size:12pt" align="center"
|style="font-weight:bold" height="35" | uL 10x buffer
| align="center" | 10
| align="center" | 10
| align="center" | 10
| align="center" | 10
| align="center" | 10
| align="center" | 10
| align="center" | 10
| align="center" | 10
|- style="font-size:12pt" align="center"
|style="font-weight:bold" height="32" | uL DpnII (50U/uL)
| align="center" | 2
| align="center" | 2
| align="center" | 2
| align="center" | 2
| align="center" | 2
| align="center" | 2
| align="center" | 2
| align="center" | 2
|- style="font-size:12pt" align="center"
|style="font-weight:bold" height="15" | uL nf H20
| align="center" | 53
| align="center" | 53
| align="center" | 53
| align="center" | 53
| align="center" | 53
| align="center" | 53
| align="center" | 53
| align="center" | 53
|- style="font-size:12pt" align="center"
|style="font-weight:bold" height="15" | Total
| align="center" | 100
| align="center" | 100
| align="center" | 100
| align="center" | 100
| align="center" | 100
| align="center" | 100
| align="center" | 100
| align="center" | 100
|}
2. Incubated 16 hours at 37C, followed by 20 minutes at 65C; Continued on [[Daniel:Notebook/HiResChrPaint/2013-5-12|5/12/2013]]

Latest revision as of 21:34, 13 May 2013

Probe Prep Test 2 (Started 5/10/2013)[edit]

Back to Calendar

Ethanol Precipitation[edit]

7. Centrifuged 10000 rpm for 30 minutes at 4C
8. Removed supernatant added 750 uL 75% EtOH
9. Centrifuged 10000 rpm for 12 minutes at 4C
10. Removed supernatant, dried in hood
11. Resuspended with 50 uL
12. Column Purification, eluted with 50 uL
13. measured concentration in nanodrop

Nanodrop Results[edit]

Sample V4A V4B V4C V4D V6A V6B V6C V6D
ng/uL 127.5 156.3 130.2 130.6 142.7 138.9 146.4 133.4
ng 5100 6252 5208 5224 5708 5556 5856 5336
ug in 4 sets       21.8 22.5

DpnII Digestion[edit]

Enzyme Info

1. Prepared samples according to table
  *DpnII-Lot 0091201; Buffer 0031105
Sample V4A V4B V4C V4D V6A V6B V6C V6D
ng/uL 127.5 156.3 130.2 130.6 142.7 138.9 146.4 133.4
ng 5100 6252 5208 5224 5708 5556 5856 5336
uL 35 35 35 35 35 35 35 35
uL 10x buffer 10 10 10 10 10 10 10 10
uL DpnII (50U/uL) 2 2 2 2 2 2 2 2
uL nf H20 53 53 53 53 53 53 53 53
Total 100 100 100 100 100 100 100 100
2. Incubated 16 hours at 37C, followed by 20 minutes at 65C; Continued on 5/12/2013